首页|炎调方通过PD-1/PD-L1通路抑制脓毒症大鼠T淋巴细胞衰竭的实验研究

炎调方通过PD-1/PD-L1通路抑制脓毒症大鼠T淋巴细胞衰竭的实验研究

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目的 研究炎调方对脓毒症大鼠T淋巴细胞及T淋巴细胞表面表面程序性死亡蛋白(PD-1)、程序性死亡蛋白配体(PD-L1)表达的影响.方法 SD大鼠(雄性)分为正常组、假手术组、模型组和炎调方组.盲肠结扎穿孔(CLP)制备脓毒症大鼠模型,于造模后 12、24、48、72 h采集外周血处死大鼠(每个时间点各 5 只大鼠),采用流式细胞分析法测定CD3+、CD4+、CD8+比例;双抗体夹心酶联免疫吸附法(ELISA)测定CD4+及CD8+表面PD-1、PD-L1 表达水平.结果 白介素-6(IL-6)在脓毒症模型组 12h分泌达到高峰后逐渐下降,降钙素原(PCT)在模型组 24h达到高峰后逐渐下降;炎调方组在 12h降低IL-6 的水平(P＜0.05),24 h时明显降低IL-6、PCT的水平(P＜0.01).模型组CD3+、CD4+、CD8+比例早期分泌开始减少,48 h分泌达最低点(P＜0.01);炎调方组在 48h时可升高 CD3+、CD4+、CD8+比例,提高CD4+/CD8+的比值而具体调节T细胞活化和增殖的功能.模型组CD4+、CD8+表面PD-1、PD-L1 表达水平早期 24h即表达增多(P＜0.01),72 h表达最多;炎调方组 24h时即可抑制PD-1、PD-L1 的表达水平而具体改善T细胞衰竭的作用.CD4+的比例与CD4+表面PD-1、PD-L1 的表达呈负相关(P＜0.01);CD8+比例与CD8+表面PD-1、PD-L1 的表达呈负相关(P＜0.01).结论 脓毒症早期 48h内即出现了T细胞活化和增殖的功能衰竭引起的T细胞免疫抑制.炎调方能在脓毒症早期改善T细胞活化和增殖的功能,其机制可能与抑制PD-1/P-L1 通路有关.
Experimental study on inhibition of T lymphocyte failure by Yantiao Formula through the PD-1/PD-L1 pathway in septic rats
Objective To study the effect of Yantiao Formula on the expression levels of programmed cell death protein-1(PD-1)and programmed cell death ligand-1(PD-L1)in T lymphocytes and on T lymphocyte surface of septic rats.Methods Male Sprague-Dawley(SD)rats were divided into control group,sham operation group,model group and Yantiao Formula group.The cecal ligation and puncture(CLP)method was adopted to prepare the sepsis model in rats.After 12 h,24 h,48 h and 72 h of modeling,rats were subjected to the collection of peripheral blood samples and sacrificed.The proportions of CD3+,CD4+and CD8+T cells were measured by flow cytometry.Expression levels of PD1 and PD-L1 on the surface of CD4+and CD8+cells were measured by enzyme-linked immunosorbent assay(ELISA).Results We detected the expression levels of inflammatory factor interleukin 6(IL-6)and infection indicator procalcitonin(PCT)in rats.In model group,IL-6 level peaked at 12 h and gradually decreased,and PCT peaked at 24 h and gradually decreased.In Yantiao Formula group,IL-6 level was reduced from 12 h(P＜0.05),and significantly reduced IL-6 and PCT levels were detected at 24h(P＜0.01).The proportions of CD3+,CD4+and CD8+T cells in the model group were reduced in the early stage,and bottomed at 48h(P＜0.01).In Yantiao Formula group,the proportions of CD3+,CD4+and CD8+T cells were elevated at 48 h.Increasing CD4+/CD8+was capable of regulating the activation and proliferation of T cells.Expression levels of PD1 and PD-L1 on the surface of CD4+and CD8+T cells were upregulated as early as at 24 h in the model group,and peaked at 72h(P＜0.01).The treatment of Yantiao Formula effectively inhibited PD1 and PD-L1 on the surface of CD4+and CD8+T cells as early as at 24 h,thus alleviating T lymphocyte failure.The proportion of CD4+T cells was negatively correlated with expression levels of PD1 and PD-L1 on the surface of CD4+T cells(P＜0.01),and the proportion of CD8+T cells was negatively correlated with expression levels of PD1 and PD-L1 on the surface of CD8+T cells(P＜0.01).Conclusion T lymphocyte failure of the suppressed activation and proliferation occurs in the early stage at 48 h of sepsis.Yantiao Formula improves T cell activation and proliferation in the early stage of sepsis by inhibiting the PD-1/PD-L1 pathway.

Yantiao Formulasepsis with T lymphocyte failureprogrammed cell death protein-1(PD-1)/programmed cell death ligand-1(PD-L1)pathway

王文清、王庆、熊旭东、方荣

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201203 上海市,上海中医药大学附属曙光医院急诊科(含重症医学科)兼感染科

炎调方 脓毒症T淋巴细胞衰竭 PD-1/PD-L1通路

国家自然科学基金面上项目国家自然科学基金面上项目上海市临床重点专科建设项目上海中医药大学附属曙光医院四明青年基金

8197453982174121shslczdzk04402SGKJ-201903

2024

10.3969/j.issn.1002-7386.2024.10.004

河北医药
河北省医学情报研究所

河北医药

CSTPCD
影响因子：1.075
ISSN：1002-7386
年,卷(期)：2024.46(10)
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